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We narrowed to 863 results for: TOR;

Showing: 321 - 340 of 863 results

  1. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ...bacteria that have taken up this vector, but it won’t tell you whether the vector contains the gRNA insert or...X-gal allows you to identify cells that took up the vector and distinguish between plasmids that contain the...functional 𝛽-galactosidase. Blue-white screens rely on vectors where the gene of interest is inserted into the..., you will see two bands: one representing your vector backbone and the other representing the insert....doesn’t contain the insert, you will just see the vector backbone band. Or, you can choose an enzyme that...specific to your insert or by using primers that are vector specific but amplify a potential insert. If you...plasmid does not contain the insert. If you’re using vector-specific primers, look for size differences. You...

  2. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    Published
    July 28, 2017, 1:30 p.m.
    ... your gene of interest in a plasmid and get the vector incorporated into the germline. To begin this process...knockdown. Addgene also has a number of insect vectors to consider. An easy way to see what is available...Find Gal4 and UAS plasmids at Addgene Choosing a vector to generate a transgenic fly There are a few important...important points to consider when choosing the vector you’d like to use to generate your new fly line:... gene to be incorporated into the genome? Some vectors incorporate into the genome randomly, while others...would you like to select for your flies? Sure, the vector will have an antibiotic resistance gene to select...transgene. One such marker is the white gene, which restores eye color to fly lines where the endogenous white...

  3. From Lab Bench to Leadership: Transitioning into Management as a PhD

    Type
    Blog Post
    Published
    Jan. 21, 2025, 2:15 p.m.
    ...project management. Mentorship is another invaluable resource: connecting with mentors who have made similar...management often means stepping into unfamiliar territory. Unlike solving technical problems in the lab,...lab, management involves a different skill set: mentoring, setting strategic goals, and fostering team success...for me. I began exploring alternative career trajectories within the industry, but the transition wasn... role is also crucial. I owe quite a bit to my mentors, as they encouraged me to explore small-scale opportunities...company by taking on responsibilities such as mentoring a junior colleague. Guiding an intern, in particular...and leveraging tools like the Myers-Briggs Type Indicator or DISC assessments can help you navigate team...

  4. Oh, The Places You Can Go: Careers in Science Communication - Writing for a Research Institute

    Type
    Blog Post
    Published
    Feb. 2, 2018, 3:20 p.m.
    ...feels that storytelling is an impactful tool for science communicators. “The power of a story is to make... to learn things quickly. Story telling. The ability to know what’s a story and how to hook your readers...there’s always plenty of writing: plain-language stories about Fred Hutch research, pieces for donors such...that can be a very painful process. And certain stories can make you feel like ‘I’m a terrible writer. ...through connections and that’s something that good storytellers in science can do.” Susan’s number one piece...social media as a scientist here Learn more about storytelling in science ...

  5. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    Published
    Jan. 29, 2015, 1:25 p.m.
    ...AMA I am a strong advocate for peer mentoring, but volunteer mentors are only able to go so far with their...coach do that is different from an adviser, boss or mentor? First, professional coaches have experience and...infrastructure are not always as effective in mentoring for this transition. Second, a coach will help...experiences. Good coaches will have loads of success stories and plenty of people to vouch for them. Trust...help you. A good coach usually will be a clear communicator and make direct suggestions for improvement ...online content for PhDs and postdocs posted by Toronto-based Jennifer Polk of From PhD to Life. She runs...has an outstanding blog and, while her PhD is in History, I think she would be a good match as a coach for...

  6. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    Published
    Dec. 1, 2015, 3:30 p.m.
    ...scientists who were potential Addgene requestors and depositors. China is in one of the top 10 countries...using bacterial phage vectors with CRISPR/cas9 sensitizing gene cassettes. Thse vectors are engineered to ...world. With three Nobel Prize winners, one NIH Director and a co-founder of BGI-China presenting, it’s... discoveries and moving beyond DNA Eric Green, Director of the National Human Genome Research Institute...human tRNA genes and, until now, we've overlooked regulatory fragments of these transcripts. I remember when...was kicked off by Nobel Laureate (and Addgene depositor) Craig Mello. Taking his groundbreaking work in...Medicine presented another unexpected finding in the regulatory RNA field. The Rajewsky lab is studying circular...

  7. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    Published
    May 2, 2024, 1:15 p.m.
    ...utility of the pRDA_526 vector. Moreover, the screen identified putative novel regulators of CD8+ T cell responses... here! Marty LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard Medical School ...in immune cells have identified many putative regulators of immunity. However, in vitro screens cannot... of this system is a lentiviral gRNA expression vector (pRDA_526) that (1) enables efficient knockout ...model in vivo. In this screen, we used the pRDA_526 vector to knock out 30 genes putatively involved in CD8...responses to cancer. The screen recovered known regulators, such as Ptpn2, Pdcd1, and Zc3h12a, demonstrating...

  8. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...into wide use, there were a variety of ways to monitor cell, organelle, and protein localization. For ...the FP timers, add an entire new dimension to monitoring localization; using FP timers, researchers can...Tsuboi et al.), limiting its use. Nonetheless, its creators noticed a few properties that have now become ...benefit of the common FP timers is that you can monitor age by expressing a single FP-timer protein or ... These separate proteins should be useful for monitoring cellular events occurring at varied time scales...fluorescent protein timers FP timers have been used to: Monitor dynamics of vesicle fusion and release from the...vesicle cargoes to FP timer mk-GO (Tsuboi et al.) Monitor gene expression in the developing pancreas by placing...

  9. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ...Administration of substances to laboratory animals: routes of administration and factors to consider. PubMed, 50...type of injection to use depends on a number of factors. You have to weigh the pros and cons of various...be injected, including cells, drugs, DNA, viral vectors, and viruses. All researchers must go through ...choice for administering different drugs and viral vectors. Like IV injections, intraperitoneal injections...common use of intradermal injections is studying inflammatory responses, as this method is useful for analyzing...for the delivery of various therapies and viral vectors. This injection sounds very disruptive, but surprisingly... This is most often used for delivery of viral vectors, though drug treatments are also possible. One ...

  10. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ...bacteria, but have no fear - plasmids from Addgene depositors are making it easier than ever to use CRISPR  ...double stranded breaks (DSB) are lethal. Addgene depositor Luciano Marraffini’s lab took advantage of this...activation of transcription including transcription factors, phage proteins, and RNAP components (Dong et al... al., 2018). They found a transcription factor called SoxS for CRISPRa. Adding complexity, they found ...been observed with Phagemids (Selle et al., 2020, Citorik et al., 2015, Bikard et al., 2014). When the CRISPR... on the presence of specific target sequences (Citorik et al., 2014). These could be genes, specific polymorphisms...refined by the addition of a uracil DNA glycosylase inhibitor and degradation tag on the fusion protein to limit...

  11. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    Published
    March 21, 2019, 12:47 p.m.
    ...different Cas12a inhibitors. Some of these anti-Cas12a proteins have broad inhibitory effects on Cas12a...Caixia Gao’s lab has developed a new plant base editor from an evolved tRNA adenosine deaminase that is... is fused to a nicking Cas9. This base editor can be used for A•T to G•C edits in protoplasts or plants... Biology Find plasmids containing these base editors Spatially and temporally regulated AAV-based ...single step gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one overnight...overnight reaction. STAgR is highly customizable as vector backbones, and gRNA structure, numbers, and promoters...Mobile-CRISPRi plasmids Identification of Cas12a inhibitors Using a bioinformatics and a screening approach...

  12. The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years

    Type
    Blog Post
    Published
    Jan. 8, 2014, 3:10 p.m.
    ...zinc finger nucleases to transcription activator-like effector nucleases (TALENs). Our most popular kit... Our first popular "kit," Andrew Fire's set of vectors for C. elegans research was our most requested ...expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're familiar with...requests grow for lentiviral packaging and expression vectors, including pMD2.G and psPAX2. Actually, these two...now have 19 different ubiquitin plasmids in the repository. 100,000 Plasmids Shipped! - By 2010, Addgene...be popular years after they were added to the repository, including beta-catenin reporter M50 Super 8x... GCaMP6 series of genetically encoded calcium indicators were the hot technology of the year in our collection...

  13. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ...DNA-binding domains based on transcription-activator-like effectors (TALEs) as anchors for genome engineering... “Transcriptional activators are easy to expand or combine with different effectors. I’d encourage people...The results are light-inducible transcriptional effectors (LITEs) designed to bind specific genes and turn...devised the plasmids now available in Addgene’s repository for use in neurons, both in culture and in living...Zhang showed they could package LITEs in viral vectors and genetically target them to specific cell populations...expression levels reliably increased in vitro by a factor of 10 to 20. A flexible and modular optogenetics...

  14. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...NHEJ). Designing an RNA editor The Zhang lab envisioned a two-component RNA editor: a Cas13 enzyme fused...after that, the lab published a paper on an RNA editor that allows C to U edits (RESCUE). We’ll walk through...translational machinery treats like guanine. This RNA editor would permit point mutations in RNA which could...nonfunctional. In their quest to build a robust RNA editor, the lab started with the Cas13 scaffold, testing...PspCas13b. Find the plasmids for the RESCUE RNA editors here! Testing and improving RNA editing from the...is too large to fit into adeno-associated viral vectors, they tested truncated ADARDDs to see if they could...edit multiple RNAs. It’s clear that CRISPR RNA editors will open new doors for scientific research - how...

  15. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    Published
    July 1, 2025, 1:15 p.m.
    ...molecules can alter the overall 260 nm reading, distorting the final concentration calculation and potentially...(Figure 1B). The A260/230 ratio can be a good indicator of protein contamination, but a number of other...contaminants alter this ratio as well (Koetsier and Cantor, 2019). The 280 nm peak is more specific to proteins...researchers traditionally use the A260/280 ratio as an indicator of protein contamination. However, this ratio ...salts, and detergents (Figure 1C-D; Koetsier and Cantor, 2019). Guanidine salts and detergents have absorbance...at concentrations as low as 10 µM (Koetsier and Cantor, 2019). As such, a sample with a good A260/280 ...the presence of RNA contamination (Koetsier and Cantor, 2019). To confirm RNA in your DNA sample, you ...

  16. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...papers are analogous to mammalian viral vectors in that the vectors are replication-deficient. Thus, a very...like for organisms like C. difficile. In 2014, Citorik et al. and Bikard et al. developed sequence-specific...antimicrobial, plasmid-based, CRISPR systems (Find the Citorik Plasmids here). In both cases, they used 1-2 CRISPR...sequences found only in certain bacteria (Figure 1). Citorik et al. targeted beta-lactam and quinolone resistance... for bacterial CRISPR targeting. Phage-derived vectors carry CRISPR RNAs (crRNAs) targeting either genomic...suggest that targeting multiple resistance/virulence factors may further reduce this likelihood. The key problem...hardest-to-kill bacteria. In using phage-derived vectors to carry CRISPR, we’re turning the tables on the...

  17. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors Read our Blog...discovered the modular format of some transcriptional activators: that the DNA binding domain (DBD) and transcriptional...of whether it interacts with a transcriptional activator (1a) or a repressor (1b). Scientists took advantage... original Y2H systems utilized the yeast Gal4 activator, bacterial LexA DBD and the lacZ reporter gene...detectable reporter signals depend on a number of factors, including protein expression and correct folding... is to use a number of different bait and prey vectors. This has been shown to be as effective as using...transcription-activating function of a eukaryotic regulatory protein. Science. 1986; 231:699-704. PubMed PMID...

  18. Which Fluorescence Microscopy Technique is Best for Me?

    Type
    Blog Post
    Published
    Oct. 10, 2017, 1:57 p.m.
    ...This post was contributed by Doug Richardson, Director of the Harvard Center for Biological Imaging and...block out of focus fluorescence from reaching a detector or restricting fluorescence to specific excitation...light passes through a pinhole before reaching the detector. The pinhole is specifically placed to only allow...allow light from the focal plane to reach the detector and all light from above or below is excluded. ...the optics of a microscope before it reaches the detector. This means that two or more objects lying within...collected and projected back to a fast, sensitive detector such as a scientific CMOS camera. If plasma membrane...higher photon flux via the more efficient camera detector and lack of a light-reducing pinhole. By collecting...

  19. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    Published
    Nov. 14, 2017, 2 p.m.
    ....com. Mentors You may have an adviser, but do you have a mentor? There is a difference. A mentor is someone...that many graduate and post-doctoral programs lack any formal or mandatory professional development training... you may need to look a bit harder to find your mentor. This may be a connection you’ve made at a conference... Alternatively, it might even be a colleague. A mentor does not necessarily need to be someone a lot older...planning, has written a great blog on choosing good mentors for scientists. Continuing professional development...popular job site webpages. Importantly, find a good mentor that can share relevant experience and resources...

  20. What's New in CRISPR - March 2020

    Type
    Blog Post
    Published
    March 10, 2020, 1:15 p.m.
    ...Cytosine base editors with minimized off-target DNA editing The original cytosine base editor, BE3, induces...deamination. They generated these editors using the BE4max architecture. The new editors reported in this study ...the paper in Nature Biotechnology Find the base editor plasmids CRISPRi in Caulobacter crescentus ...ctrA, a gene encoding an essential cell cycle regulator. The lab expressed both dCas9 and the ctrA sgRNA...lab developed new fluorescently labeled piggyBac vectors to deliver uniform and sustained expression of ...editing by TET1 by co-delivering TET1 and protein factors for linking methylcytosine oxidation to DNA repair...
Showing: 321 - 340 of 863 results